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Transcriptional profiling of genes involved in embryogenic, non-embryogenic calluses and somatic embryogenesis of Valencia sweet orange by SSH-based microarray

机译:基于SSH的微阵列对瓦伦西亚甜橙胚性,非胚性愈伤组织和体细胞胚发生相关基因的转录谱分析

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Somatic embryogenesis (SE) is a most promising technology that is used for in vitro germplasm conservation and genetic improvement via biotechnological approaches in citrus. Herein, three suppression subtractive hybridization (SSH) libraries were constructed using calluses of Citrus sinensis cv. ‘Valencia’ to explore the molecular mechanisms that underlie the SE in citrus. A total of 880 unisequences were identified by microarray screening based on these three SSH libraries. Gene ontology analysis of the differentially expressed genes indicated that nucleolus associated regulation and biogenesis processes, hormone signal transduction, and stress factors might be involved in SE. Transcription factors might also play an important role. LEC1/B3 domain regulatory network genes (LEC1, L1L, FUS3, ABI3, and ABI5) were isolated in citrus SE. Some new transcription factors associated with citrus SE, like a B3 domain containing gene and HB4, were identified. To understand the influence of these isolated genes on SE competence, their expression profiles were compared among callus lines of seven citrus cultivars with different SE competence. The expression dynamics suggested that these genes could be necessary for the SE initiation and might play a role in embryogenic competence maintenance in different cultivars. On the basis of gene expression profiles, an overview of major physiological and biosynthesis processes at different developmental stages during citrus SE is presented. For the first time, these data provide a global resource for transcriptional events important for SE in citrus, and the specific genes offer new information for further investigation on citrus SE maintenance and development.
机译:体细胞胚发生(SE)是一种最有前途的技术,可通过柑橘中的生物技术方法用于体外种质保存和遗传改良。在本文中,使用柑桔(Citrus sinensis)cv的愈伤组织构建了三个抑制消减杂交(SSH)文库。瓦伦西亚(Valencia)探索构成柑橘SE的分子机制。通过基于这三个SSH库的微阵列筛选,总共鉴定出880个不等位基因。对差异表达基因的基因本体分析表明,核仁相关的调节和生物发生过程,激素信号转导和应激因素可能与SE有关。转录因子也可能起重要作用。 LEC1 / B3域调节网络基因(LEC1,L1L,FUS3,ABI3和ABI5)在柑橘SE中分离。鉴定了一些与柑橘SE相关的新转录因子,例如含有基因和HB4的B3结构域。为了了解这些分离的基因对SE能力的影响,比较了7种具有不同SE能力的柑橘品种的愈伤组织中它们的表达谱。表达动力学表明,这些基因可能是SE启动所必需的,并且可能在不同品种的胚发生能力维持中发挥作用。根据基因表达谱,概述了柑橘SE发育过程中不同发育阶段的主要生理和生物合成过程。这些数据首次为柑橘中SE的重要转录事件提供了全球资源,而特定基因为进一步研究柑橘SE的维持和发育提供了新的信息。

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